Figures from 30 of Scotland’s 32 local authorities revealed Glasgow City Council had the highest bill for supply teachers at £16.6m. North Lanarkshire Council’s expenses were the second highest at £11.5m.
The data, obtained by the Scottish Liberal Democrats through a Freedom of Information request, showed £81.5m was spent across Scotland on supply teachers in 2016/17.
Shetland Islands Council had the smallest supply teacher cost of those responding at £312,861.
West Lothian Council and North Ayrshire Council did not provide figures.
Last year data from 27 out of 32 councils showed a supply teacher cost of £66.1m in 2015/16.
The Liberal Democrats also revealed there were 795 teaching posts vacant when pupils returned to school after the summer holidays this year.
The party’s education spokesman Tavish Scott said: “Local authorities are spending more than £220,000 a day on supply teachers.
“This money could be better invested in ensuring schools have enough permanent staff in the first place. “There are many quality, hard-working supply teachers who do a great job filling classroom gaps.
“Children’s education is better when they have consistency in the classroom.
“The lack of a permanent teacher over a prolonged period can be a hindrance to their education.
“Continuity helps learning.”
Mr Scott added: “This sizeable bill for supply teachers shows it is time we gave the teaching profession a well-overdue boost. “A new wide-ranging independent review, McCrone Two, needs to look at teachers’ pay, conditions, numbers and support and pave the way for meaningful changes.
“This is the only way the government can rebuild the shattered trust between them and teachers.”
Scottish Conservative leader Ruth Davidson has attacked the Government over its failure to set up a flagship scheme announced last year that was meant to help attract graduates into teaching.
Education secretary John Swinney last week conceded the government’s targets for new teachers entering teacher training was short by about 200 people.
Mr Swinney told Holyrood’s education committee that more teachers had left the profession this year than the government had predicted. Only 3,861 new teachers are enrolled on courses in Scotland this year.